Biologically produced and metal-organic framework delivered dual-cut CRISPR/Cas9 system for efficient gene editing and sensitized cancer therapy

Yu, Jiantao1; Tang, Mao2; Zhou, Zhengdong2; Wei, Zixiang1; Wan, Feiyan1; Hou, Shengxin1; Li, Qing1; Li, Yan2; Tian, Leilei1

2024-04-01

10.1016/j.actbio.2024.02.030

ACTA BIOMATERIALIA   影响因子和分区

1742-7061

1878-7568

Manipulation of the lactate metabolism is an efficient way for cancer treatment given its involvement in cancer development, metastasis, and immune escape. However, most of the inhibitors of lactate transport carriers suffer from poor specificity. Herein, we use the CRISPR/Cas9 system to precisely downregulate the monocarboxylate carrier 1 (MCT1) expression. To avoid the self-repairing during the gene editing process, a dual-Cas9 ribonucleoproteins (duRNPs) system is generated using the biological fermentation method and delivered into cells by the zeolitic imidazolate framework-8 (ZIF -8) nanoparticles, enabling precise removal of a specific DNA fragment from the genome. For efficient cancer therapy, a specific glucose transporter 1 inhibitor (BAY -876) is co-delivered with the duRNPs, forming BAY/duRNPs@ZIF-8 nanoparticle. ZIF -8 nanoparticles can deliver the duRNPs into cells within 1 h, which efficiently downregulates the MCT1 expression, and prohibits lactate influx. Through simultaneous inhibition of the lactate and glucose influx, BAY/duRNPs@ZIF-8 prohibits ATP generation, arrests cell cycle, inhibits cell proliferation, and finally induces cellular apoptosis both in vitro and in vivo . Consequently, we demonstrate that the biologically produced duRNPs delivered into cells by the nonviral ZIF -8 carrier have expanded the CRISPR/Cas gene editing toolbox and elevated the gene editing efficiency, which will promote biological studies and clinical applications. Statement of significance The CRISPR/Cas9 system, widely used as an efficient gene editing tool, faces a challenge due to cells' ability to self-repair. To address this issue, a strategy involving dual -cutting of the genome DNA has been designed and implemented. This strategy utilizes biologically produced dual-ribonucleoproteins delivered by a metal-organic framework. The effectiveness of this dual -cut CRISPR-Cas9 system has been demonstrated through a therapeutic approach targeting the simultaneous inhibition of lactate and glucose influx in cancer cells. The utilization of the dual -cut gene editing strategy has provided valuable insights into gene editing and expanded the toolbox of the CRISPR/Cas-based gene editing system. It has the potential to enable more efficient and precise manipulation of specific protein expression in the future. (c) 2024 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

CRISPR/Cas9 Biological fermentation Anti-self-repairing dual-cut Metal-organic framework MCT1

SCI ; EI

英语

第一 ; 通讯

National Natural Science Foundation of China[51973089] ; Shen- zhen Fundamental Research Programs["JCYJ20220530115204011","JCYJ20220530113612027"] ; Shenzhen Science and Technology In- novation Commission[KQTD20170810111314625]

Engineering ; Materials Science

Engineering, Biomedical ; Materials Science, Biomaterials

WOS:001221306600001

ELSEVIER SCI LTD

Web of Science

1.Southern Univ Sci & Technol, Dept Mat Sci & Engn, 1088 Xueyuan Blvd,Nanshan Dist, Shenzhen 518055, Guangdong, Peoples R China
2.Southern Univ Sci & Technol, Dept Biol, 1088 Xueyuan Blvd,Nanshan Dist, Shenzhen 518055, Guangdong, Peoples R China

Yu, Jiantao,Tang, Mao,Zhou, Zhengdong,et al. Biologically produced and metal-organic framework delivered dual-cut CRISPR/Cas9 system for efficient gene editing and sensitized cancer therapy[J]. ACTA BIOMATERIALIA,2024,178.

Yu, Jiantao.,Tang, Mao.,Zhou, Zhengdong.,Wei, Zixiang.,Wan, Feiyan.,...&Tian, Leilei.(2024).Biologically produced and metal-organic framework delivered dual-cut CRISPR/Cas9 system for efficient gene editing and sensitized cancer therapy.ACTA BIOMATERIALIA,178.

Yu, Jiantao,et al."Biologically produced and metal-organic framework delivered dual-cut CRISPR/Cas9 system for efficient gene editing and sensitized cancer therapy".ACTA BIOMATERIALIA 178(2024).